Process Scale Purification of Antibodies

Promoting a continued and much-needed renaissance in biopharmaceutical manufacturing, this book covers the different strategies and assembles top-tier technology experts to address the challenges of antibody purification. Updates existing topics and adds new ones that include purification of antibodies produced in novel production systems, novel separation technologies, novel antibody formats and alternative scaffolds, and strategies for ton-scale manufacturing Presents new and updated discussions of different purification technologies, focusing on how they can address the capacity crunch in antibody purification * Emphasizes antibodies and innovative chromatography methods for processing

Autorentext

Uwe Gottschalk, PhD,is Chief Technology Officer at Lonza Pharma/Biotech, Switzerland. Previously, he served as Group Vice President at Sartorius Stedim Biotech (2004-2014) and in various development and manufacturing capacities at Bayer Health Care (19912004). Dr. Gottschalk received a doctorate in chemistry from the University of Münster (Germany) for work on antibody-drug conjugates at the Cancer Research Campaign Laboratories in Nottingham (UK). He is Head Lecturer at the University of Duisburg-Essen, Germany, and has written extensively in the areas of industrial biotechnology and somatic gene therapy.

Klappentext
Stay on the cutting edge of antibody purification with the insights of industry experts Biopharmaceutical companies are under extreme pressure to develop robust and economical large-scale processes while consistently meeting regulatory standards for safety and purity. Unfortunately, while upstream productivity has increased in line with demand, downstream processing is mired in a technology crisis resulting in production bottlenecks. Antibody purification is sandwiched between fermentation and drug formulation, two areas that have traditionally received the lion's share of industry awareness as well as generous research funding and attention in the literature. As a result, improvements in purification technology are needed urgently to meet the challenges of increasing demands without sacrificing product quality. The second edition of Process Scale Purification of Antibodies continues the legacy of its predecessor to offer scientists, researchers, and engineers an essential resource focused on best practices in manufacturing antibody drugs. It covers different antibody purification strategies, including new developments since the publication of the first edition, and an assembly of top-tier experts in these disparate technologies with the common aim of addressing problems in process-scale antibody purification. Recognized for providing a fresh perspective on traditional chromatography as well as alternative technologies and unit operations, the first edition covered innovative developments in protein purification like smart membranes, bioaffinity mimetics as well as orthogonal, and integrated strategies. The revision updates these topics and also introduces platform concepts that have gained traction over the ensuing yearsincluding the purification of antibodies produced in novel production systems, novel separation technologies, novel antibody formats and alternative scaffolds, and strategies for ton-scale manufacturing. Process Scale Purification of Antibodies, therefore, consolidates the unique niche occupied by its predecessor and maintains interest in this neglected subject area by promoting the continued, much-needed renaissance in antibody purification.

Inhalt
Preface xxiii

List of Contributors xxvii

**1 Downstream Processing of Monoclonal Antibodies: Current Practices and Future Opportunities 1
**Brian Kelley

1.1 Introduction 1

1.2 A Brief History of Current Good Manufacturing Process mAb and Intravenous Immunoglobulin Purification 2

1.3 Current Approaches in Purification Process Development: Impact of Platform Processes 4

1.4 Typical Unit Operations and Processing Alternatives 7

1.5 VLS Processes: TonScale Production and Beyond 10

1.6 Process Validation 12

1.7 Product Life Cycle Management 13

1.8 Future Opportunities 16

1.9 Conclusions 18

Acknowledgments 19

References 19

**2 The Development of Antibody Purification Technologies 23
**John Curling

2.1 Introduction 23

2.2 Purification of Antibodies by Chromatography Before Protein A 25

2.3 Antibody Purification After 1975 28

2.4 Additional Technologies for Antibody Purification 31

2.5 Purification of mAbs Approved in North America and Europe 34

2.6 Current Antibody Process Technology Developments 40

Acknowledgments 45

References 46

**3 Harvest and Recovery of Monoclonal Antibodies: Cell Removal and Clarification 55
**Abhinav A. Shukla and Eric Suda

3.1 Introduction 55

3.2 Centrifugation 59

3.3 Microfiltration 62

3.4 Depth Filtration 67

3.5 Flocculation 70

3.6 Absolute Filtration 71

3.7 Expanded Bed Adsorption Chromatography 73

3.8 Harvesting in SingleUse Manufacturing 74

3.9 Comparison of Harvest and Clarification Unit Operations 74

References 76

**4 NextGeneration Clarification Technologies for the Downstream Processing of Antibodies 81
**Nripen Singh and Srinivas Chollangi

4.1 Introduction 81

4.2 Impurity Profiles in Cell Cultures 83

4.3 Precipitation 84

4.4 Affinity Precipitation 89

4.5 Flocculation 90

4.6 Toxicity of Flocculants and Precipitants and Their Residual Clearance 96

4.7 Depth Filtration 97

4.8 Considerations for the Implementation of New Clarification Technologies 102

4.9 Conclusions and Future Perspectives 103

Acknowledgments 104

References 104

**5 Protein ABased Affinity Chromatography 113
**Suresh Vunnum, Ganesh Vedantham and Brian Hubbard

5.1 Introduction 113

5.2 Properties of Protein A and Commercially Available Protein A Resins 114

5.3 Protein A Chromatography Step Development 118

5.4 Additional Considerations During Development and ScaleUp 123

5.5 Virus Removal/Inactivation 127

5.6 Validation and Robustness 128

5.7 Conclusions 129

Acknowledgment 130

References 130

**6 Purification of Human Monoclonal Antibodies: NonProtein A Strategies 135
**Alahari Arunakumari and Jue Wang

6.1 Introduction 135

6.2 Integrated Process Design for Human Monoclonal Antibody Production 136

6.3 Purification Process Designs for HuMabs 136

6.4 Conclusions 149

Acknowledgments 151

References 152

**7 Hydrophobic Interaction Chromatography for the Purification of Antibodies 155
**Judith Vajda and Egbert Muller

7.1 Introduction 155

7.2 HIC With mAbs 156

7.3 HIC with Membrane Adsorbers 173

7.4 Future Perspectives 174

References 175

**8 Purification of Monoclonal Antibodies by MixedMode Chromatography 181
**Pete Gagnon

8.1 Introduction 181

8.2 A Brief History 182

8.3 Prerequisites for Industrial Implementation 183

8.4 Mechanisms, Screening, and Method Development 185 8.5 Ca...